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US 2013/0157266 A1Published application
Abscription based molecular detection of dna methylation
AssigneeRibomed Biotechnologies, Inc.
InventorsMichelle M. Hanna, David McCarthy
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Abstract
The present invention provides methods for detecting biomarkers based on Abscription®, abortive transcription technology. Particularly, the present invention provides bisulfate free methods for detecting methylation of CpG islands from small samples containing DNA, including formalin fixed, paraffin embedded samples. The methods are suitable for multiplexing and can be used to analyze multiple CpG islands from a single sample in a short time.
Claims18
A method for detecting methylation of a target polynucleotide in a sample comprising:
- a) cleaving a genomic DNA sample containing at least one methylated target polynucleotide with a restriction enzyme that does not cleave the methylated target polynucleotide;
- b) contacting the cleaved genomic DNA with an immobilized MBD, thereby immobilizing methylated genomic DNA from the sample;
- c) optionally, recovering the methylated genomic DNA from the immobilized MBD, thereby isolating methylated genomic DNA fragments;
- d) contacting the methylated DNA with a primer pair that specifically hybridizes to and amplifies a target sequence of the at least one polynucleotide;
- and
- e) detecting the amplified polynucleotide.
3 more elements
Dependencies are inferred from the claim text.